Assessment of clinical validity of KPG index for 3D classification of impacted maxillary canines by cone beam computed tomography in patients.

OBJECTIVES: The primary objective of this study was to assess the validity of the KPG index in predicting the difficulty of treatment involving impacted maxillary canines. The secondary objective was to assess the reliability and reproducibility of the index. MATERIALS AND METHODS: A retrospective study was conducted on 96 maxillary impacted canines (MIC) in 60 patients aged 13-35 years. Cone-beam computed tomography (CBCT) scans were used to predict the treatment difficulty of MIC using the KPG index. Patient case files were assessed for the actual difficulty encountered in treating MIC. Cohen's kappa correlation coefficient was used for intra-observer reliability and Kendell's W test was used for inter-observer reliability. Spearman's correlation coefficient test was used to assess the correlation between predicted and actual treatment. RESULTS: Easy and moderately difficult cases exhibited a moderate correlation between actual and predicted treatment outcomes, whereas difficult cases displayed a weak correlation. The perfect correlation was observed exclusively in extremely difficult cases. The intra-observer reliability for assessing CBCT scans using the KPG guide was found to be 0.88, and the inter-rater reliability was 0.94. CONCLUSION: The KPG index displayed 87%, 71%, 50% and 100% validity in easy, moderately difficult, difficult, and extremely difficult cases, respectively. This index showed good reliability and reproducibility. However, it is imperative to consider a multitude of other factors, including the patient's age, presence of associated root resorption in adjacent teeth, and duration of treatment, to make an informed decision between surgical exposure and extraction.

Culture-independent real-time PCR reveals extensive polymicrobial infections in hospitalized diarrhoea cases in Kolkata, India.

Culture-independent identification of diarrhoeal aetiological agents was performed using DNA harvested from diarrhoeal stool specimens with SYBR-Green-based real-time PCR targeting Vibrio cholerae, Vibrio parahaemolyticus, Campylobacter spp., Shigella spp. and three different pathotypes of diarrhoeagenic Escherichia coli. Conventional culture-dependent methods detected bacterial enteropathogens in 68 of 122 diarrhoeal stool specimens. Of 68 specimens, 59 (86.8%) had a single pathogen and the remaining nine (13.2%) had polymicrobial infections with multiple pathogens. Re-analysis of the 68 specimens by culture-independent real-time PCR methods showed that 25 (36.8%) specimens contained single pathogen and 43 (63.2%) specimens contained mixed infections with multiple pathogens. The prevalence of such high levels of polymicrobial infections would not have been detected without using real-time PCR. Culture-dependent analysis assigned 54 of the 122 selected archived specimens as 'no known aetiology'. However, re-analysis of these samples by real-time PCR showed the presence of single or multiple pathogens among 34 (63%) of these specimens. Estimation of relative pathogen load by real-time PCR in the stool specimens indicated that the inability of conventional culture-dependent methods to detect the pathogens was related to lower colony-forming units of the pathogen, as reflected by lower C(t) values. Detection of high levels of polymicrobial infection by real-time PCR indicates that in the settings like Kolkata and its surroundings, where cholera and other enteric diseases are endemic, the concept of one pathogen one disease might need to be re-evaluated.

An easy protocol for studying chromatin and recombination protein dynamics during Arabidopsis thaliana meiosis: immunodetection of cohesins, histones and MLH1.

Plant meiosis studies have enjoyed a fantastic boom in recent years with the use of Arabidopsis thaliana as a model not only for molecular genetics and genomics but also for cytogenetics. In this article we describe a new protocol for immunolabelling meiotic proteins that allows the detection of a large range of proteins on strongly spread chromosomes throughout the entire meiotic process. We used this method to immunodetect MLH1, a crucial component of the meiotic recombination machinery, and found that it can be visualised as foci from pachytene to diakinesis, where it co-localises with chiasmata. The mean MLH1 foci number per meiotic cell at diakinesis was 8.4 for WS-4 and 9.95 for Col-0, with the number of foci per bivalent ranging from 1 to 5. We also analysed MLH1 distribution within bivalents and found that they were not restricted to specific chromosomal regions. The analysis of MLH1 foci formation in the Atzip4 mutant, where class I crossover (CO) formation is prevented, revealed that residual chiasmata were not labelled by MLH1, strongly suggesting that MLH1 antibodies only label class I COs in Arabidopsis. It thus appears that the 'obligatory CO' is systematically labeled by MLH1 and is generated through the class I pathway.

RAD51 loss of function abolishes gene targeting and de-represses illegitimate integration in the moss Physcomitrella patens.

Gene targeting (GT) is a major tool for basic and applied research during which the transforming DNA, which shares sequence homology with a chromosomal target, integrates at the corresponding locus by homologous recombination (HR). In eukaryotes, GT recruits enzymes from the HR-mediated double strand break repair pathway. Different mechanisms of HR have been described which depend on the Rad52 epistasis group of genes, but which specific mechanism is used by the cell for GT remains unclear. In Saccharomyces cerevisiae, the RAD52 protein is essential for GT, and the RAD51 protein plays a minor role. In filamentous fungi and animal cells, however, GT depends on RAD51 and is weakly affected by suppression of RAD52. Genetic evidence also indicates that the non-homologous end-joining pathway of DSB repair has a negative impact on GT efficiencies, but how the balance between these two pathways is controlled is poorly understood. Here, we have examined the role of RAD51 in the only plant that exhibits high GT frequencies, the model bryophyte Physcomitrella patens. Our results show that the two RAD51 proteins have partially redundant functions in the maintenance of genome integrity and resistance to ionizing radiation. Furthermore, we demonstrate that loss of function of the two RAD51 proteins completely abolishes GT and strongly increases illegitimate integration rates in this moss. These findings demonstrate for the first time in plant the critical role of RAD51 in controlling the balance between targeted and random integration events observed upon transgenesis, and confirm that P. patens is a particularly interesting tool for studying GT in higher eukaryotes.

Thyroid hormone induces the generation of a novel putative protein in piscine ovarian follicle that stimulates the conversion of pregnenolone to progesterone.

Ovarian follicles were collected from the perch belonging to the vitellogenic stage and incubated in vitro for 4 h in the absence (control) or presence of triiodothyronine (T3). Addition of T3 (40 ng/ml) to the follicle incubation caused a two-fold increase of [3H] pregnenolone conversion to radiolabelled progesterone (P4) as compared to the control. The increase in P4 formation in the ovarian follicle could be blocked completely by the inhibitors of protein synthesis, actinomycin D and cycloheximide (50 micrograms/ml), suggesting a protein or peptide mediator of the T3 stimulatory effect. To search for this mediator, ovarian follicles from the control or T3 incubate were homogenized and ultracentrifuged and different fractions were added separately to fresh follicle incubations. Only the 100,000 g supernatant from T3 incubate showed a significant (p < 0.01) increase in P4 formation, while the corresponding supernatant from control follicle incubations had no such stimulatory effect. Trypsin or heat destroyed this augmentory effect. Based on its ability to stimulate the conversion of radiolabelled pregnenolone to P4 in the ovarian follicle, the T3-induced protein (TIP) was purified to homogeneity by employing Sephadex G-75 gel filtration, FPLC Mono-Q and FPLC Superose-6 chromatography. Sodium dodecyl sulphate polyacrylamide gel electrophoresis of purified TIP showed it to be a 52 K monomer protein. Addition of TIP in increasing concentrations to follicle incubations caused a linear increase in P4 formation. Experiments with radiolabelled TIP ([125I] TIP) indicate its entry through the follicular cell membrane within the limited period of incubation. Results suggest that TIP activates ovarian 3 beta-hydroxysteroid dehydrogenase enzyme, thus effecting a greater conversion of pregnenolone to P4.

CNS practice in neurosurgery.

The roles and responsibilities of a neurosurgical clinical nurse specialist are described in relation to the five traditional roles (clinician, educator, consultant, coordinator, and researcher). These roles are referred to in relation to a CNS practicing in an expanded role in a neurosurgery department. The primary role and major impact of the CNS are to augment patient care delivery through communication and coordination of physician and nursing services. This augmentation is achieved through accuracy, efficiency, and effectiveness of the CNS functioning in all five roles.